Wang Xiaoying, Peng Jiefu, Wang Jing, Li Miao, Wu Di, Wu Songyan, Liao Jipei, Dou Jun
Department of Basis Medicine, Wuxi School of Medicine, Jiangnan University, Wuxi, People's Republic of China.
Department of Pathogenic Biology and Immunology, School of Medicine, Southeast University, Nanjing, People's Republic of China.
Onco Targets Ther. 2018 Mar 2;11:1173-1182. doi: 10.2147/OTT.S149254. eCollection 2018.
Despite the breadth of understanding the noncoding RNAs' function in molecular biology, their functional roles in hepatocellular carcinoma (HCC) is poorly understood. In this study, we investigated the effect of hepatitis C virus (HCV) core upon the expression of noncoding RNAs.
The lncRNAs, mRNAs, and circRNAs were employed for identification of HCV core protein gene expression in human Huh7 hepatoma (Huh7) cell line. In data analysis, we applied a threshold that eliminated all genes that were not increased or decreased by at least a 2-fold change in a comparison between transfected and control cells. Hierarchical Clustering and the Kyoto encyclopedia of genes and genome pathway analyses were performed to show the distinguishable lncRNA, mRNAs, and circRNAs expression pattern among samples.
The array data showed that 4,851 lncRNAs, 4,785 mRNAs, and 823 circRNAs were 2-fold up-regulated but 3,569 lncRNAs, 3,192 mRNAs, and 419 circRNAs were 2-fold down-regulated in Huh 7-core cells. The genes in the enriched set were associated with macromolecule and nucleic acid metabolic processes, DNA damage response and regulation of voltage-gated calcium channel. We identified 10 genes from the selected 14 genes that were higher or lower expression in Huh7-core cells than that of Huh7-vector cells by quantitative real-time polymerase chain reaction. Interestingly, overexpression of miR122 and miR204 partly abrogated the expression of TGFBRAP1 and HOTTIP, and increased the HPCAL1 expression in the predicted carcinogenic pathways.
Our data suggests that the pathways of miR204-HPCAL1-lncRNAHOTTIP and miR122-TGFBRAP1 were likely involved in the carcinogenic progress due to the presence of HCV core, and that overexpression of miR122 and miR204 might inhibit the HCC progress by down-regulation of TGFBRAP1 and HOTTIP expression.
尽管对非编码RNA在分子生物学中的功能已有广泛了解，但其在肝细胞癌（HCC）中的功能作用仍知之甚少。在本研究中，我们调查了丙型肝炎病毒（HCV）核心蛋白对非编码RNA表达的影响。
使用长链非编码RNA（lncRNA）、信使核糖核酸（mRNA）和环状RNA（circRNA）来鉴定人Huh7肝癌细胞系中HCV核心蛋白基因的表达。在数据分析中，我们应用了一个阈值，去除了在转染细胞与对照细胞的比较中，表达未至少增加或减少2倍的所有基因。进行层次聚类分析和京都基因与基因组百科全书通路分析，以显示样本间lncRNA、mRNA和circRNA的不同表达模式。
阵列数据显示，在Huh 7-核心蛋白细胞中，4851个lncRNA、4785个mRNA和823个circRNA上调了2倍，但3569个lncRNA、3192个mRNA和419个circRNA下调了2倍。富集集中的基因与大分子和核酸代谢过程、DNA损伤反应以及电压门控钙通道调节有关。通过定量实时聚合酶链反应，我们从选定的14个基因中鉴定出10个基因，其在Huh7-核心蛋白细胞中的表达高于或低于Huh7-载体细胞。有趣的是，在预测的致癌途径中，miR122和miR204的过表达部分消除了TGFBRAP1和HOTTIP基因的表达，并增加了HPCAL1的表达。
我们的数据表明，由于HCV核心蛋白的存在，miR204-HPCAL1-lncRNAHOTTIP和miR122-TGFBRAP1通路可能参与致癌过程，并且miR122和miR204的过表达可能通过下调TGFBRAP1和HOTTIP的表达来抑制HCC进展。
